PURIFICATION OF CELL-POPULATIONS FROM HUMAN FETAL BRAIN USING FLOW CYTOMETRIC TECHNIQUES

被引:11
作者
ROZENTAL, R
GEBHARD, D
PADIN, C
URBAN, M
WU, JY
SPRAY, DC
CHIU, FC
机构
[1] UNIV FED RIO DE JANEIRO, INST BIOPHYS CARLOS CHAGAS FILHO, BR-21941 RIO DE JANEIRO, BRAZIL
[2] ALBERT EINSTEIN COLL MED, DEPT MICROBIOL & IMMUNOL, NEW YORK, NY USA
[3] ALBERT EINSTEIN COLL MED, SAUL KOREY DEPT NEUROL, NEW YORK, NY USA
[4] UNIV KANSAS, DEPT PHYSIOL & CELL BIOL, KANSAS CITY, KS USA
[5] ALBERT EINSTEIN COLL MED, DEPT MED, NEW YORK, NY USA
来源
DEVELOPMENTAL BRAIN RESEARCH | 1995年 / 85卷 / 02期
关键词
FLOW CYTOMETRY; GLUTAMIC ACID DECARBOXYLASE; NEUROFILAMENT PROTEIN; HUMAN NEURON;
D O I
10.1016/0165-3806(94)00204-D
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
We recently established primary cultures from dissociated second trimester human fetal brains using a novel spin seeding method and characterized cellular populations with distinct phenotypes in these cultures. Here, we report that these neural cultures can be dissociated to single-cell suspensions, sorted by size using flow cytometry and re-seeded to yield cultures selectively enriched for the neuronal and glial cell populations. Sorted neurons were highly homogeneous, viable and extended processes by one day after re-seeding. These neurons expressed immunoreactivity for neurofilament protein, retained their GABAergic phenotype and were electrically excitable. Re-seeded astrocytes proliferated in culture and expressed glial fibrillary acidic protein. We describe the conditions required for the flow cytometric sorting and tissue culture assays as well as the morphological, immunocytochemical and electrophysiological characteristics of the sorted neuronal population.
引用
收藏
页码:161 / 170
页数:10
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